Cartilage production promoter and prophylactic or therapeutic agent for diseases associated with cartilage damage

ABSTRACT

Disclosed is a therapy which is for preventing or treating cartilage damage and diseases associated with cartilage damage, such as arthritides and osteoarthritis, and utilizes a more effective and more safe medicinal agent. Specifically disclosed are a cartilage production promoter, a glucosaminoglycan and/or proteoglycan production promoter, and a prophylactic or therapeutic agent for diseases associated with cartilage damage, each of which comprises fucoidan as an active ingredient.

TECHNICAL FIELD Related Application

This application claims the benefit of the priority of Japanese PatentApplication No. 2009-73856 filed to the Japanese Patent Office on Mar.25, 2009. The patent application to which this application claimspriority is incorporated into the present specification by reference inits entirety.

The present invention relates to a chondrogenesis promoter, aglycosaminoglycan and/or proteoglycan production promoter, and aprophylactic or therapeutic agent for a cartilage damage and a diseasedue to a cartilage damage, which contain a fucoidan as an effectivecomponent.

BACKGROUND ART

Cartilage is a tissue composed of a cartilage matrix and cartilage cells(chondrocytes) spotted therein. The cartilage matrix is composed ofwater, collagen, proteoglycan, and others. Proteoglycan is a complexcomposed of a polysaccharide called glycosaminoglycan, and a protein.Cartilage fulfills various functions in the living body through theelasticity thereof.

Diseases due to a cartilage damage include various diseases such asarthrosis deformans, arthritis and rheumatism. As therapeutic agents forthese diseases, many effective components such as glucosamine (seeNon-Patent Document 1) and glucuronic acid (Patent Document 1) areknown.

Fucoidan is a polysaccharide contained in marine alga (for example,mozuku (Nemacystus decipiens), Japanese kelp (Laminaria japonica) etc.),and has been ingested from early times by eating of these marine alga.Fucoidan has many physiological activity functions such asimmunostimulatory activity, adhesion-preventing activity andanti-inflammatory activity (see Patent Documents 2 and 3, and others).

Patent Document 4 describes that fucoidan promotes a morphogeneticaction of bone and cartilage. However, the disclosure of Patent Document4 is merely about an agent for promotion of osteogenesis. Osteoblast isnecessary to induce bone, and chondroblast is necessary to inducecartilage. Osteoblast and chondroblast are each derived bydifferentiation or induction from undifferentiated cells in accordancewith different systems. Osteoblast and chondroblast are entirelydifferent cells, and are also different from each other in theirproperties and functions. Osteoblast produces type I collagen, andcalcifies it with alkaline phosphatase activity, so as to ossify.Chondroblast, which is cartilage cell precursor, produces type IIcollagen or glycosaminoglycan and is differentiation-induced tocartilage cell. The cartilage cell synthesizes proteoglycan around thecells, thereby blocking calcification and bone formation based onosteoblast to form cartilage. For the formation of cartilage,ossification is required to be inhibited. The cartilage formation is apromoting of synthesis of proteoglycan by cartilage cell. Thus thecartilage formation is a biological phenomenon which is entirelydifferent from the bone formation. In other words, Patent Document 4discloses a reverse effect to a matter that fucoidan promotes theformation of cartilage.

In Examples of Patent Document 4, results obtained by administeringfucoidan or the like directly into established cell lines are disclosed.However, according to any ordinary administering manner such as oralingestion, fucoidan is never directly contacted with osteoblast. Inconclusion, Patent Document 4 never discloses what effect is producedwhen fucoidan is administered into an actual living body.

As described above, there has not been any finding that fucoidan itselfhas an effect of promoting a chondrogenesis.

-   Patent Document 1: JP-A-2004-10533-   Patent Document 2: JP-A-2005-82806-   Patent Document 3: JP-A-2005-508893-   Patent Document 4: JP-A-2004-10533-   Non-Patent Document 1: Tamai et al., Carbohydrate Polymers, 2002,    vol. 48, pp. 369-378

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention

Many patients suffer from arthropathy, and further it is anticipatedthat patients of senile arthrosis will increase hereafter as thepopulation ages in Japan. In such a situation, there are still desiresfor more useful therapeutic agents capable of repairing a cartilagedamage. It should be preferred that such therapeutic agents are safe andfree from any side effects, and can prevent diseases when routinelyingested.

Means for Solving the Problems

The present inventors have intensively studied to solve the problems,and a result, have found out fucoidan as a substance for solving theproblems. Thus, the present invention has been completed.

That is, the present invention provides:

(1) A chondrogenesis promoter comprising a fucoidan as an effectivecomponent,

(2) The chondrogenesis promoter according to (1), wherein the fucoidanhas a molecular weight of 8,000 to 400,000,

(3) The chondrogenesis promoter according to (1), wherein the fucoidanhas a molecular weight of 50,000 to 150,000,

(4) A glycosaminoglycan and/or proteoglycan production promotercomprising a fucoidan as an effective component,

(5) The glycosaminoglycan and/or proteoglycan production promoteraccording to (2), wherein the fucoidan has a molecular weight of 8,000to 400,000,

(6) The glycosaminoglycan and/or proteoglycan production promoteraccording to (2), wherein the fucoidan has a molecular weight of 50,000to 150,000,

(7) A pharmaceutical or veterinary medical agent composition forpreventing or treating a cartilage damage and a disease due to acartilage damage, comprising a fucoidan as an effective component,

(8) The pharmaceutical or veterinary medical agent composition accordingto (7), wherein the fucoidan has a molecular weight of 8,000 to 400,000,

(9) The pharmaceutical or veterinary medical agent composition accordingto (7), wherein the fucoidan has a molecular weight of 50,000 to150,000,

(10) Use of a fucoidan in a production of a pharmaceutical or veterinarymedical agent composition for preventing or treating a cartilage damageand a disease due to a cartilage damage,

(11) The use according to (10), wherein the fucoidan has a molecularweight of 8,000 to 400,000,

(12) The use according to (10), wherein the fucoidan has a molecularweight of 50,000 to 150,000,

(13) A method for preventing or treating a cartilage damage or a diseasedue to a cartilage damage, which comprises administering a fucoidan inan effective amount to an animal,

(14) The method according to (13), wherein the fucoidan has a molecularweight of 8,000 to 400,000, and

(15) The method according to (13), wherein the fucoidan has a molecularweight of 50,000 to 150,000.

Effects of the Invention

According to the present invention, a chondrogenesis promoter, aglycosaminoglycan and/or proteoglycan production promoter, and aprophylactic or therapeutic agent for a cartilage damage and a diseasedue to a cartilage damage, which contain a fucoidan as an effectivecomponent, are obtained. These are useful for preventing and treating,for example, arthrosis deformans, senile arthrosis, and articularrheumatism. Since these contain a fucoidan, which is a natural substancecontained in marine algae, these are high in safety and give noside-effect.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows visual evaluation and comparison of the extent of healingof damages in the control group and the fucoidan-administered group.

FIG. 2 shows results obtained by staining proximal trochlear grooves ofthe control group and those of the fucoidan-administered group withalcian blue. The numbers 1, 2 and 3 show the rabbit individuals in therespective groups; and the upper shows the control group, and the lowershows the fucoidan-administered group.

FIG. 3 shows results obtained by staining distal trochlear grooves ofthe control group and those of the fucoidan-administered group withalcian blue. The numbers 1, 2 and 3 show the rabbit individuals in therespective groups; and the upper shows the control group, and the lowershows the fucoidan-administered group.

FIG. 4 shows results obtained by staining proximal trochlear grooves ofthe control group and those of the fucoidan-administered group withsafranin-O. The numbers 1, 2 and 3 show the rabbit individuals in therespective groups; and the upper shows the control group, and the lowershows the fucoidan-administered group.

FIG. 5 shows results obtained by staining distal trochlear grooves ofthe control group and those of the fucoidan-administered group withsafranin-O. The numbers 1, 2 and 3 show the rabbit individuals in therespective groups; and the upper shows the control group, and the lowershows the fucoidan-administered group.

FIG. 6 shows results obtained by analyzing alcian blue-stained imagesand safranin-O-stained images of damaged site samples.

FIG. 7 shows results obtained by analyzing alcian blue-stained imagesand safranine-O-stained images about a control group, and damaged sitesamples in which fucoidans having a molecular weight of 239,000, and330,000 were each administered.

FIG. 8 shows visual evaluation and comparison of the extent of healingof damages in a control group, and fucoidan-administered groups in whichfucoidans having different molecular weights were each administered.

FIG. 9 shows results obtained by analyzing alcian blue-stained images ofdamaged site samples of a control group, and fucoidan-administeredgroups in which fucoidans having different molecular weight were eachadministered.

FIG. 10 shows results obtained by analyzing safranine-O-stained imagesof damaged site samples of a control group, and fucoidan-administeredgroups in which fucoidans having different molecular weight were eachadministered.

FIG. 11 shows results obtained by staining damaged sites of a controlgroup and groups of 8,000 and 50,000 in molecular weight with alcianblue. Each image in the lower in the view shows an enlarged image of arectangular region in the corresponding image in the upper.

FIG. 12 shows results obtained by staining damaged sites of groups of146,000, 239,000, and 330,000 in molecular weight with alcian blue. Eachimage in the lower in the view shows an enlarged image of a rectangularregion in the corresponding image in the upper.

FIG. 13 shows results obtained by staining damaged sites of groups of400,000, and 1,000,000 in molecular weight with alcian blue. Each imagein the lower in the view shows an enlarged image of a rectangular regionin the corresponding image in the upper.

BEST MODE FOR CARRYING OUT THE INVENTION

The present inventors have found out that fucoidan has a function ofpromoting a chondrogenesis, and repairing a damage of cartilage. Thepresent inventors have further found out that fucoidan promotes theproduction of glycosaminoglycan and proteoglycan. Fucoidan is apolysaccharide contained in marine algae (for example, mozuku(Nemacystus decipiens) and Japanese kelp (Laminaria japonica) etc.), andhas been ingested from early times by the eating of these marine alga.Fucoidan has many physiological activity functions such asimmunostimulatory activity, adhesion-preventing activity, andanti-inflammatory activity. However, there has not been any finding thatfucoidan itself has an effect of promoting the chondrogenesis.

In the first aspect, the present invention provides a chondrogenesispromoter containing a fucoidan as an effective component.

In the second aspect, the present invention provides a glycosaminoglycanand/or proteoglycan production promoter containing a fucoidan as aneffective component.

In the third aspect, the present invention provides a prophylactic ortherapeutic agent for a cartilage damage and a disease due to acartilage damage, containing a fucoidan as an effective component.

The chondrogenesis promoter, the glycosaminoglycan and/or proteoglycanproduction promoter and the prophylactic or therapeutic agent for acartilage damage and a disease due to a cartilage damage, of the presentinvention, are useful for preventing or treating symptoms related to acartilage damage, for example, arthrosis deformans, senile arthrosis,articular rheumatism, spondylitis deformans, articular sprain, andligament damage.

The fucoidan used as an effective component in the chondrogenesispromoter, the glycosaminoglycan and/or proteoglycan production promoterand the prophylactic or therapeutic agent thereof for a cartilage damageand a disease due to a cartilage damage, of the present invention, canbe a purified product, or a roughly purified product, for example, anextract from marine algae such as mozuku. The method or means forextracting fucoidan from fucoidan-containing materials such as marinealgae is well known. The fucoidan used in the present invention can bein the form of a solid (such as powder and granule), a liquid (such asaqueous solution of fucoidan, fucoidan suspension and extract frommarine algae), or a semi-solid (such as paste). In the presentinvention, a fucoidan-containing material can be contained as aneffective component. Preferred examples of the fucoidan-containingmaterial include marine algae, in particular brown algae. Examples offucoidan-containing brown algae include mozuku (for example,okinawamozuku (Cladosiphon okamuranus) and itomozuku (Nemacystusdecipiens)), wakame (Undaria pinnatifida), mekabu (sporophyl of wakame),arame (Eisenia bicyclis), gagome (Kjellmaniella crassifolia), Japanesekelp (Laminaria japonica), kurome (Eckronia kurome), kajime(Kjellmaniella crassifalia), mitsuishikonbu (Laminaria angustata),yoremoku (Sargassum siliquastrum), hiziki (Hizikia fusiformis),hondawara (Sargassum fulvellum), yatsumatamoku (Sargassum patens),akamoku (Sargassum horneri), Arctic wrack (Focus evanescens), andumitoranoo (Sargassum thunbergii). However, the invention is not limitedby the examples.

The chondrogenesis promoter, the glycosaminoglycan and/or proteoglycanproduction promoter, and the prophylactic or therapeutic agent for acartilage damage and a disease due to a cartilage damage, of the presentinvention, can be in the form of a pharmaceutical composition. Themethod for administering the composition is not particularly limited,and can be any method such as an oral, injection, or percutaneousmethod. An oral administration is preferred. In the case of an oraladministration agent, the composition can be formulated into variousoral agent forms such as condensate, powder, granule, tablet, capsuleagent and drinking agent. Methods for producing these agent forms arewell known, and the following processes can be appropriately used:mixing, dissolving, pulverizing, tableting, drying, and others. Acarrier, or excipient can be used in accordance with the purpose of thepharmaceutical composition. A flavor, a sweetener, a colorant and/orsome other can be appropriately added to any oral administration agentof the present invention. The oral administration agent of the presentinvention can be administered as it is, or can be administered in thestate that the agent is appropriately added to a food or drink such asmiso soup and tea since the fucoidan as an effective component has notaste and smell.

The chondrogenesis promoter, the glycosaminoglycan and/or proteoglycanproduction promoter, and the prophylactic or therapeutic agent for acartilage damage and a disease due to a cartilage damage, of the presentinvention, can be in the form of a food or drink. Since fucoidan itselfhas no taste and smell, fucoidan is used to produce various foods ordrinks without producing any effect onto flavor and taste. For example,miso soup or soup can be produced into which fucoidan powder, or anfucoidan extract from mozuku or some other is incorporated.Alternatively, the promoter or agent of the present invention can bemade into a form obtained by concentrating a fucoidan extract frommozuku or some other and packaging the concentrate into a package, orsubjecting the extract to freeze-drying or some other treatment to makethe concentrate into a powder or granular form or some other form andthen putting the resultant into an appropriate packaging or container,or by subjecting the extract to some other processing, and thereby,users themselves can add the promoter or agent of the present inventionto any foods or drinks to ingest.

The chondrogenesis promoter, the glycosaminoglycan and/or proteoglycanproduction promoter, and the prophylactic or therapeutic agent for acartilage damage and a disease due to a cartilage damage, of the presentinvention, can be a supplement containing a fucoidan or afucoidan-containing material. The supplement can be made into the formof tablet, capsule agent, granule, powder, or some other by a methodwell known by those skilled in the art. As described herein, when thefood or drink according to the present invention is used, a user or apatient can ingest the fucoidan routinely over a long term withouthesitation. Thus, the promoter or agent of the present invention cancontribute to the prevention and the treatment of the above-mentioneddiseases. The food or drink according to the present invention is usefulalso as a functional food.

A dose of the chondrogenesis promoter, the glycosaminoglycan and/orproteoglycan production promoter, and the prophylactic or therapeuticagent for a cartilage damage and a disease due to a cartilage damage, ofthe present invention, can be decided by those skilled in the artwithout difficulty. For example, the amount of the fucoidan to beadministrated can be decided while the preventing or treating effectagainst a target disease such as arthrosis deformans, senile arthrosisand articular rheumatism is observed. The amount (dry amount) of thefucoidan to be administered through the agent of the present inventionis usually from about 0.3 g or more per day, preferably about 1 g ormore per day for an adult.

The chondrogenesis promoter, the glycosaminoglycan and/or proteoglycanproduction promoter, and the prophylactic or therapeutic agent for acartilage damage and a disease due to a cartilage damage, of the presentinvention, can contain one or more effective components other thanfucoidan. Examples of the other effective component include knownanti-inflammatory agents, and other therapeutic agents (for example,glucosamine etc.) for diseases due to a cartilage damage.

Furthermore, the present invention provides the use of a fucoidan in theproduction of a chondrogenesis promoter, a glycosaminoglycan and/orproteoglycan production promoter, and a prophylactic or therapeuticagent for a cartilage damage and a disease due to a cartilage damage.

In another aspect, the present invention provides a veterinary medicalagent, for preventing or treating a cartilage damage and a disease dueto a cartilage damage, that contains a fucoidan as an effectivecomponent. The veterinary medical agent can be in the form of aveterinary medical agent composition. The method for administrating theveterinary medical agent is not particularly limited, and can be anymethod such as oral, injection and percutaneous method. Oraladministration is preferred. In the case of an oral administrationagent, the composition can be formulated into various oral agent forms,such as condensate, powder, granule, tablet, capsule agent and drinkingagent. Methods for producing these agent forms are well known, and thefollowing processes can be appropriately used: mixing, dissolving,pulverizing, tableting, drying, and others. A carrier, or excipientacceptable for veterinary medical agents can be used. A flavor, asweetener, a colorant and/or some other can be appropriately added toany oral administration agent of the present invention. The veterinarymedical agent can contain one or more other effective components otherthan fucoidan. Examples of the other effective component include knownanti-inflammatory agents and therapeutic agents (for example,glucosamine etc.) for diseases due to a cartilage damage.

The present invention further provides the use of a fucoidan in theproduction of a veterinary medical agent for preventing or treating acartilage damage and a disease due to a cartilage damage.

The present invention further provides a method for preventing ortreating a cartilage damage and a disease due to a cartilage damage in atarget animal which includes a human or does not include human, in whicha fucoidan is administered in an effective amount to the animal.

The molecular weight of the fucoidan as an effective component in thepresent invention is preferably in the range of 8,000 to 1,000,000, forexample, 8,000 to 250,000, 250,000 to 500,000, 500,000 to 750,000,750,000 to 1,000,000, 8,000 to 350,000, 350,000 to 1,000,000, 50,000 to1,000,000, 150,000 to 1,000,000, 250,000 to 1,000,000, 400,000 to1,000,000 or the like. The molecular weight is more preferably in therange of 8,000 to 40,000, for example, 8,000 to 150,000, 150,000 to250,000, 250,000 to 350,000, 350,000 to 400,000, 50,000 to 350,000,50,000 to 400,000, 150,000 to 350,000, 150,000 to 400,000, 250,000 to400,000 or the like. The molecular weight is even more preferably in therange of 8,000 to 250,000, for example, 8,000 to 50,000, 50,000 to100,000, 100,000 to 200,000, 8,000 to 150,000, 150,000 to 250,000,50,000 to 250,000 or the like. The molecular weight is furtherpreferably in the range of 50,000 to 150,000, for example, 50,000 to100,000, 100,000 to 150,000 or the like. The molecular weight of 150,000is most preferably.

The molecular weight of the fucoidan can be measured by a well knownmethod, for example, gel filtration chromatography. Usually, pullulanhaving an already known molecular weight is used as a marker when themolecular weight is measured. Generally, the molecular weight ofpolymeric compound is represented by the average molecular weight, thenumber-average molecular weight or the weight-average molecular weight.In the specification, the term “molecular weight” denotes weight-averagemolecular weight unless otherwise specified. The weight-averagemolecular weight can be calculated by a well known means or method, andfor example, can be calculated from the pattern of gel filtrationchromatography.

When any molecular weight is represented by a numerical value in thespecification, the molecular weight include values in the range of about±20% of the numerical value. For example, in the case of a molecularweight of 50,000, this molecular weight includes molecular weight valuesof about 40,000 to about 60,000.

Hereinafter, the present invention will be specifically described inmore detail by way of examples. However, the present invention is notlimited by the examples.

EXAMPLE 1 Example 1 Production of Damaged Models

As animal specimens, groups of female white rabbits (weight: about 2.0kg) were used, the number of the rabbits in each of the groups being 3.The rabbits were habituated to the environment for one week from thetime of delivery. Thereafter, 0.1 mg/kg of medetomidine wassubcutaneously injected into each of the rabbits, and 25 mg/kg ofKetalar was intramuscularly injected thereto. About each of the rabbitspecimens subjected to the injection anesthesia, the left knee joint wasshaved, and then disinfected. Thereafter, approaching from the outsideof the knee joint, the articular capsule was opened and the knee jointwas exposed completely by shifting the patella to the inside. Withreference to the experiment of Tamai et al. (Carbohydrate Polymers,2002, vol. 48, pp. 369-378), three holes of 2 mm in diameter and 4 mm indepth were bored with a hand-drill at the proximal trochlear groove,distal trochlear groove, and the medial trochlear ridge. Thereafter, thesites were sufficiently rinsed, and then the articular capsule wassutured with 3-0PDS (polydioxanone). The subcutaneous tissue and theskin were simultaneously sutured with stainless steel wires. After theoperation, 0.5 mg/kg of atipamezole was diluted and subcutaneouslyinjected two times per day to wake the rabbits.

From the day of the operation, initially every day, water (50 mL) inwhich 1 g of a fucoidan was dissolved were dosed to the rabbits in thefucoidan-administered group by natural intake. Thereafter, water wasgiven to the rabbits to drink freely. This was continued for 3 weeks.Only water was given to the rabbits in the control group. During thisperiod, there were no signs of side effects or the like in thefucoidan-administered group.

After the end of the fucoidan-administering term of the three weeks, therabbit specimens in each of the fucoidan-administered group and thecontrol group were euthanized, and then the operated site was exposed.

EXAMPLE 2 Example 2 Macroscopic Observation of the Degree of Healing

The operated site of the left femur in each of the damaged models inExample 1 was macroscopically observed. About the degree of healingafter the 3 weeks, many of the damages were satisfactorily cured in thefucoidan-administered group. In the control group, many of the damageswere not cured. The degree of healing of each of the damages was scoredon the basis of Table 1, and evaluated.

TABLE 1 Remark through macroscopic observation Score Healing rate ofless than 50% 0 Healing rate of 50% or more and less 1 than 60% Healingrate of 60% or more and less 2 than 80% Healing rate of 80% or more 3

The results were as shown in FIG. 1. From the results, in thefucoidan-administered group, good therapeutic results were obtained inthe proximal trochlear groove and the distal trochlear groove. In thecontrol group, the damaged sites were filled with collagen fibers,fibroblasts and others. By contrast, in the fucoidan-administered group,the damaged sites were substituted with chondroblasts and cartilagetissues.

EXAMPLE 3 Histological Observation

The left femur of each of the damaged models in Example 1 was recovered,and then fixed with a 10% neutral buffered formalin aqueous solution(formaldehyde solution). The tissue was then decalcified with a 5%formic acid solution under shaking. After the decalcification, thetissue was neutralized with a 5% sodium sulfate solution, and washedwith water and then dehydrated. The tissue piece in which thedecalcification had been completed was cut so that the repaired siteswere positioned on a vertical cross section. Thereafter, the cut tissuewas embedded with paraffin in the usual manner, and then sliced with amicrotome into thin pieces in thickness of 5 μm. The resultant tissueswere used and each stained with alcian blue (AB) and safranin-O (SO) toprepare samples for histological observation. The samples were observedwith a microscope, and subjected to image analysis. About the imageanalysis, a 200-power image of each of the repaired sites was digitizedby use of Photograb ab-300 version 1.0 (Macintosh Software, AdobeSystems Inc., Tokyo). Pixels, the number of which was 20,000, were setat random in 6 positions of the image. The number of pixels in which atarget color tone occupied was measured. The obtained numerical valuewas subjected to statistical processing (t-test).

(a) Observed Results of the Staining with Alcian Blue

FIGS. 2 and 3 show results obtained by observing, with the microscope,the proximal and distal trochlear groove in the tissue samples stainedwith alcian blue. In the fucoidan-administered group, the repairedregions were stained into blue (densely colored regions in the figures).Thus, it showed that a large amount of glycosaminoglycan was contained.By contrast, stained regions were hardly observed in the control group.

(b) Observed Results of the Staining with Safranin-O

FIGS. 4 and 5 show results obtained by observing, with the microscope,the proximal and distal trochlear groove in the tissue samples stainedwith safranin-O. In the fucoidan-administered group, the repairedregions were stained (densely colored regions in the figures). Thus, itshowed that a large amount of proteoglycan was contained. By contrast,stained regions were hardly observed in the control group.

(c) Results of the Image Analysis

FIG. 6 shows results obtained by analyzing the images stained withalcian blue and the images stained with safranin-O by theabove-mentioned method. According to the results, thefucoidan-administered group exhibited a significantly higher value abouteach of the two stains than the control group. Thus, it showed thatglycosaminoglycan and proteoglycan were present in a significantlylarger amount in the fucoidan-administered group.

According to the results of Examples 2 and 3, it is considered that, inthe fucoidan-administered group, the production of glycosaminoglycan andproteoglycan were promoted in the cartilage tissues and that the repairof the cartilage tissues was promoted.

EXAMPLE 4 Example 4 Comparison in the Effects between Fucoidans having aMolecular Weight of 239,000 and 330,000

In the production of the same damaged models as in Example 1, fucoidanshaving a molecular weight of 239,000 and 330,000 were each administeredin fucoidan-administered groups. Each of the administered groups weresubjected to staining with alcian blue and safranin-O, and imageanalysis, according to the method of Example 3.

FIG. 7 shows results about the groups in which the fucoidan having thedifferent molecular weights were each administered and the controlgroup. The results were obtained by analyzing the images stained withalcian blue and the images stained with safranin-O by theabove-mentioned method. According to the results, in each of thefucoidan-administered groups, the production of glycosaminoglycan andproteoglycan were promoted. In the group in which thesmaller-molecular-weight, which was 239,000, fucoidan was administered,glycosaminoglycan and proteoglycan were each produced in a largeramount.

EXAMPLE 5 Example 5 Comparison 1 in the Effects between Fucoidans havingMolecular Weights Different from Each Other; Production of DamagedModels

In the production of the damaged models of Example 1, damaged modelgroups in which fucoidans having a molecular weight of 8,000, 50,000,146,000, 239,000, 330,000, 400,000, and 1,000,000 were each administeredwere produced.

EXAMPLE 6 Example 6 Comparison 2 in the Effects between Fucoidans havingMolecular Weights Different from Each Other; Macroscopic Observation ofthe Degree of Healing

The operated sites of the left femurs in each of the damaged modelgroups in Example 5 were macroscopically observed. About each of thedamaged model groups, the degree of healing was evaluated on the basisof the criterion in Example 2. The obtained numerical values weresubjected to a multiple comparison test according to Turkey-Kramermethod. The results were as shown in FIG. 8. From the results, all ofthe fucoidan-administered groups were better in therapeutic results thanthe control group. In particular, the groups of 50,000, 146,000, and400,000 in molecular weight showed better results.

EXAMPLE 7 Example 7 Comparison 3 in the Effects between Fucoidans havingMolecular Weights Different from Each Other; Histological Observation

Each of the damaged model groups in Example 5 was histologicallyobserved according to the method in Example 3. The obtained numericalvalues were subjected to a multiple comparison test according toTurkey-Kramer method. The results of the image analysis are shown inFIGS. 9 and 10. From the results, all of the fucoidan-administeredgroups were better in therapeutic results than the control group. Inparticular, the groups of 50,000, 146,000, and 239,000 in molecularweight showed better results. The group of 146,000 in molecular weightshowed the best results.

FIGS. 11 to 13 show microscopically observed results of the alcianblue-stained samples of the control group, and each of the damaged modelgroups. White dots observed in the fucoidan-administered groupsrepresent chondroblasts. As shown by enlarged images in the lower, itwas observed that chondroblasts were activated in each of the damagedmodels. In the group 146,000 in molecular weight, which showed the bestresults, it was observed that chondroblasts were further activated.

INDUSTRIAL APPLICABILITY

The present invention is directed to a chondrogenesis promoter, aglycosaminoglycan and/or proteoglycan production promoter, and aprophylactic or therapeutic agent for a cartilage damage and a diseasedue to a cartilage damage, which contain a fucoidan as an effectivecomponent. The present invention provides, for example, a pharmaceuticalcomposition or a food or drink. Thus, the present invention is usable inthe field of pharmaceuticals, foods and drinks, and others.

1. A chondrogenesis promoter comprising a fucoidan as an effectivecomponent.
 2. The chondrogenesis promoter according to claim 1, whereinthe fucoidan has a molecular weight of 8,000 to 400,000.
 3. Thechondrogenesis promoter according to claim 1, wherein the fucoidan has amolecular weight of 50,000 to 150,000.
 4. A glycosaminoglycan and/orproteoglycan production promoter comprising a fucoidan as an effectivecomponent.
 5. The glycosaminoglycan and/or proteoglycan productionpromoter according to claim 2, wherein the fucoidan has a molecularweight of 8,000 to 400,000.
 6. The glycosaminoglycan and/or proteoglycanproduction promoter according to claim 2, wherein the fucoidan has amolecular weight of 50,000 to 150,000.
 7. A pharmaceutical or veterinarymedical agent composition for preventing or treating a cartilage damageand a disease due to a cartilage damage, comprising a fucoidan as aneffective component.
 8. The pharmaceutical or veterinary medical agentcomposition according to claim 7, wherein the fucoidan has a molecularweight of 8,000 to 400,000.
 9. The pharmaceutical or veterinary medicalagent composition according to claim 7, wherein the fucoidan has amolecular weight of 50,000 to 150,000.
 10. Use of a fucoidan in aproduction of a pharmaceutical or veterinary medical agent compositionfor preventing or treating a cartilage damage and a disease due to acartilage damage.
 11. The use according to claim 10, wherein thefucoidan has a molecular weight of 8,000 to 400,000.
 12. The useaccording to claim 10, wherein the fucoidan has a molecular weight of50,000 to 150,000.
 13. A method for preventing or treating a cartilagedamage or a disease due to a cartilage damage, which comprisesadministering a fucoidan in an effective amount to an animal.
 14. Themethod according to claim 13, wherein the fucoidan has a molecularweight of 8,000 to 400,000.
 15. The method according to claim 13,wherein the fucoidan has a molecular weight of 50,000 to 150,000.